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Updated: Jun 2, 2026

A New Screening Method for the Directed Evolution of Thermostable Bacteriolytic Enzymes
Published on: November 7, 2012
Dramatically improved catalytic activity of an artificial (S)-selective arylmalonate decarboxylase by
Yusuke Miyauchi1, Robert Kourist, Daisuke Uemura
1Department of Biosciences and Informatics, Keio University, 3-14-1 Hiyoshi, Kohoku-ku, Yokohama, Kanagawa, 223-8522, Japan.
Structure-guided directed evolution significantly enhanced arylmalonate decarboxylase activity. This resulted in a 220-fold increase in (S)-naproxen production with high enantioselectivity.
Area of Science:
- Biocatalysis
- Enzyme Engineering
Background:
- Arylmalonate decarboxylase (ADC) is crucial for synthesizing chiral compounds.
- Improving ADC catalytic efficiency and enantioselectivity is vital for pharmaceutical production.
Purpose of the Study:
- To enhance the catalytic activity and enantioselectivity of an (S)-selective arylmalonate decarboxylase variant.
- To optimize the production of (S)-naproxen using the engineered enzyme.
Main Methods:
- Employed three rounds of structure-guided directed evolution.
- Utilized G74C/C188S variant of arylmalonate decarboxylase.
Main Results:
- Achieved up to a 920-fold increase in catalytic activity.
- The best variant demonstrated a 220-fold improved activity for (S)-naproxen production.
- Maintained excellent enantioselectivity (>99% ee).
Conclusions:
- Structure-guided directed evolution is effective for enzyme optimization.
- Engineered ADC variants show significant potential for industrial chiral synthesis.
- High-yield and enantioselective production of (S)-naproxen is achievable.
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